LOCUS       sequence-211521-        7864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7864)
  TITLE     yeast expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTAU-CN-DM
FEATURES             Location/Qualifiers
     source          1..7864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(143..162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(203..219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(203..219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(216..238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    227..243
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(251..257,258..275,276..281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    296..317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        411..626
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             627..1430
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(1635..1652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2714..2733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3393..4184,4185..4253))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4016..4035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4254..4358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4426..4444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4482..4504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4604..4623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4832..4848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5138..5158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5186..5310
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(5739..6044)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(5841..5860)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6364..7047)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     6962..6981
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7048..7150)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(7328..7452)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      7469..7716
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7485..7503)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7734..7753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7735..7753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7763..7779)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
       61 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      181 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      361 cgtgccagca gatctgagca gattgtactg agagtgcacc ataccacctt ttcaattcat
      421 catttttttt ttattctttt ttttgatttc ggtttccttg aaattttttt gattcggtaa
      481 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
      541 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
      601 aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
      661 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
      721 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
      781 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
      841 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
      901 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
      961 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1021 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1081 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     1141 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     1201 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     1261 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     1321 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     1381 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     1441 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     1501 tattacccta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1561 aaagatctgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1681 gtatcagcat cgatgctaga gtcgaatgga aattacgtac tttcaacgca agagcaagac
     1741 aaaagaaagc ctaaaactgt cttttcgtgt cggtcgtcca atatcatcgt aaacgtgtgt
     1801 agagccttaa attctaattt ctaactttaa attttttttg ctattataat actaatttct
     1861 aactttcaac ctattttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt
     1921 tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt
     1981 atagcacgtg atgaaaagaa atatagtctt caaggttata ttatacaggt ttatatatta
     2041 ttttacttat ttttgttgtt ctttttcttg gagtatttag cattaacaac ttcgacccat
     2101 cgcgatgacg aaattgcacg tttaaaggag aagtcggaat cgttgcaaga agaaataaat
     2161 tagtttgacg ttttattcct tttcttctgc acagaagtgt gtatatatat atttatatat
     2221 agtcgatgtg caaaagccta cattattttt cttttgattt ttattttttt gttcaagtag
     2281 gttcatgccg acagtaaatc cttatgccct acctaaatat aaacacccaa atatatatat
     2341 atatatatat atatataaat aataaaaacg gaaaggaaat agaataataa taataatact
     2401 acacgtttac gtatgtaact cattctatta ttcagacttt gatattaata gaattattac
     2461 tactaggttt cctcacttca aaatctataa atataccgtc atgctctaat cgatgctcac
     2521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4381 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4441 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4501 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4561 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4621 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4681 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4741 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     4801 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     4861 cttttgctgg catttcttct agaagcaaaa agagcgatgc gtcttttccg ctgaaccgtt
     4921 ccagcaaaaa agactaccaa cgcaatatgg attgtcagaa tcatataaaa gagaagcaaa
     4981 taactccttg tcttgtatca attgcattat aatatcttct tgttagtgca atatcatata
     5041 gaagtcatcg aaatagatat taagaaaaac aaactgtaca atcaatcaat catcacataa
     5101 aatggggtcg aggaaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga
     5161 attgggtacc gcgtcgacgg atctcacaag tttgtacaaa aaagctgaac gagaaacgta
     5221 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     5281 ctgtaaaaca caacatatcc agtcactatg aatcaactac ttagatggta ttagtgacct
     5341 gtagtcgacc gacagccttc caaatgttct tcgggtgatg ctgccaactt agtcgaccga
     5401 cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc cagcctactc gctattgtcc
     5461 tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga ggtgcgagcc tcttttttgt
     5521 gtgacaaaat aaaaacatct acctattcat atacgctagt gtcatagtcc tgaaaatcat
     5581 ctgcatcaag aacaatttca caactcttat acttttctct tacaagtcgt tcggcttcat
     5641 ctggattttc agcctctata cttactaaac gtgataaagt ttctgtaatt tctactgtat
     5701 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     5761 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     5821 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     5881 tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg
     5941 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     6001 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     6061 ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     6121 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     6181 taccagaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     6241 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     6301 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     6361 cttttagtaa gccggatcca cgcggcgttt acgccccccc tgccactcat cgcagtactg
     6421 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     6481 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     6541 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     6601 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     6661 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     6721 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     6781 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     6841 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     6901 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     6961 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     7021 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     7081 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     7141 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     7201 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     7261 aagtgcgtcg ggtgatgctg ccaacttagt cgactacagg tcactaatac catctaagta
     7321 gttgattcat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     7381 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     7441 gtacaaagtg gtaagtaact aactcgagtc atgtaattag ttatgtcacg cttacattca
     7501 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     7561 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     7621 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     7681 gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata
     7741 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     7801 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     7861 gcga
//
